Antibody Purification Resins—A Better Alternative to Protein A

Antibody purification using thiophilic resins is a simple, powerful, and economical alternative to Protein A, the most common method of immunoglobulin purification. Although Protein A antibody purification is very common, there are certain types of antibodies, such as the single-chain antibodies IgE, IgY, and IgM, that cannot be purified using Protein A. An alternative antibody purification method, thiophilic affinity chromatography (TAC), is ideal for these types of applications, as well as immunoglobulin purification in general, including IgG, IgM, IgA, Fab and Fc fragments, and C3 and C4 complement factors.

TAC is a powerful and simple method for the purification of proteins—in particular, immunoglobulins. It is an economical alternative to Protein A for purifying antibodies from whole serum and tissue culture. In comparison to Protein A-based immunoadsorbents, thiophilic adsorbents have a broader affinity for immunoglobulins. Further, more than 99% of total proteins are recovered using a thiophilic adsorbent, in comparison to less than 92% for phenyl and 75% for octyl agarose adsorbents.

TAC has broad specificity for immunoglobulins of different classes and subclasses from a wide range of species and has capacity similar to that of other affinity methods.

Its advantages are the simplicity of the sample preparation, applicability with a range of source materials and protein concentration ranges, and gentle elution conditions, unlike the popular Protein A/Protein-G separation methods that require extreme pH for elution, which the antibodies may not be stable enough to accomodate.

Features

Thiophilic-Superflow Resin utilizes thiophilic adsorption chromatography. With this technique, protein adsorbs to a sulfone thioether ligand. The adsorption of different proteins can be promoted by adding different salts to the mixture. Varying the concentration of the loading salt can affect the adsorption affinities of IgG, IgM, IgA, Fab and Fc fragments, and C3 and C4 complement factors.

Purification with Thiophilic-Superflow Resin offers several advantages over purification with Protein A, the conventional method used for immunoglobulin purification:

Purification at neutral pH

Broader selectivity (IgY, IgM, IgE, scAb)

High linear flow rates

High capacity (15–20 mg Ab/ml resin)

Reusability

Higher stability of the purified product

Applications

Antibody purification, including IgG, IgM and chicken IgY

Properties of Thiophilic-Superflow Resin

Feature

Thiophilic-Superflow Resin

Batch/gravity

Yes

FPLC

Yes

Scale

Analytical, preparative

Preparative production capacity (mg IgG/ml absorbant)

25

Matrix

Cross-linked agarose

Maximum linear flow rate (cm/min)

5.0

Maximum volumetric flow rate (ml/min) at 5 x 1 cm i.d.

4.0

pH stability

2–10

Supplied as

Bulk/slurry 50% in 25% ethanol

Storage

4°C, do not freeze

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.